Enteric bacteria boost defences against oxidative stress in Entamoeba

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Entamoeba histolytica. Hugo Varet, Yana Shaulov, ... of E. histolytica to NS. Trophozoites (1x106) were treated with (i) GSNO (350 uM) for 1 hour at 37°C (NI);.
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Enteric bacteria boost defences against oxidative stress in Entamoeba histolytica Hugo Varet, Yana Shaulov, Odile Sismeiro, Meirav Trebicz-Geffen, Rachel Legendre, Jean-Yves Coppée, Serge Ankri and Nancy Guillen Scientific Reports, 2018 _______________________________________________________________________________________________ Included material Figure S1: Effect of Escherichia coli O55 on the resistance of E. histolytica to nitrosative stress. Figure S2: Effect of diverse bacteria on the resistance of E. histolytica to oxidative stress. Figure S3: GO Terms enrichment in E. histolytica under stress upon incubation with diverse bacteria. Document DOC1: LRR motifs in the amoebic proteins identified in this work (see Table S8).

Figure S1. Effect of Escherichia coli O55 on the resistance of E. histolytica to NS. Trophozoites (1×106) were treated with (i) GSNO (350 μM) for 1 hour at 37°C (NI); (ii) 1×109 live E. coli O55 (LB) for 30 minutes and then with GSNO (350 μM) for 1 hour; and (iii) heat-killed E. coli O55 (KB, the equivalent of 1×109 bacteria) for 30 minutes and then with GSNO (350 μM) for 1 hour. The trophozoites’ viability was determined in an eosin dye exclusion assay.





Figure S2. Effect of diverse bacteria on the resistance of E. histolytica to oxidative stress

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Trophozoites (1×10 ) in the presence of the diverse live bacteria (1×10 ) were treated with 2.5 mM H2O2 for 1 hour at 37°C. The trophozoites’ viability was determined in an eosin dye exclusion assay.

Figure S3. GO Terms enrichment in E. histolytica under stress upon incubation with diverse bacteria.

The data from Supplementary Table 6 were analysed with PANTHER tools (http://pantherdb.org/), in order to determine the fold enrichment (FE) of gene sets with GO terms, relative to the background list for all genes in the amoeba genome. The GO terms were used to identify enriched molecular processes; those with an FE ≥ 2.5 were plotted.

Supplementary DOC 1. LRR motifs in the amoebic proteins identified in this work (see Table S8). The diverse LRR motifs were localized by visual inspection of amino acid sequences. We took into account the consensus sequences for LRR Bsp A: LxxIxLxxxVxxIgxxAFxxCxx LRR11: LxxLxLxxNxL LRR12: LxxLxLxxCxxL EHI_120570, 356 aa MQLGYNEIMIVSKYFDDINDFINLEIGVKRFQGNMERFHFNPIPLNQYSRKLFPNIETF HIYNIYDEVFEDGRIFKYVIWYKVDYSRYLEEKEEMNEYKNIEYTQEDRKKYGNTIPNE VTSLGYWCFSLCDDIQEIEIPTSVTEIRSYCFERCSS LQTITIPTNVSKIGDSCFYKCSS LQTITIPTNVSKIRECCFYGCSS LTTINIPPSVTKIGNRCFEECYS LTSINIED-VKYISEKRIFMNEPV LISIKILENLTIINGKNIEKKDINEFIIPSSITKLGDYCFEYCYS LQTINIPTSVSKIGDYCFKECTS LTSINIPSSITSFGKGCFYHCRCEEELKKNKTIPEYCFEE EHI_139980, 666 aa MQRDNPNNMKRIRRKMMSLNIPTRPFLLTENTGSIIPTLSEIILDSFGRFEQNFSSQQE QTPIALPPVLASKLIYRISNNPNASESDLNIILFRILSSGTPLQVIDLHHTHLLTKFTC MNLSQLCNNVSITTLNISYATGISSESLKVLFRSFKQLKYLNTEACVGFDDNAFISLLT IHPPLEVIVISNCPKISDKSVIELKNINSLISFKANNIQLTISSISVLHGLKEIELMGN NCLDDQCLLKISQLNPDLTRICFGNSRLSDEALQRLLQQLNGNLISLNLSGCTRAGPMT LAQLFNSQFKLKYLNLANCFGINGDSILESYHPMFRSTLFRWLEEIEYLNISGCIRLSE EFITTTLIASPQLRSLILDDTAVTDQCLQEFIQSSITYNQTVRRSIHTPGMKVHPLTLS LKRCIRLTDNAYTNLFQTRGVDLISLNLSFCNSLSSLSLNVFSLYSNTLTSFYASNNDM ISEYSWLKFIASCPQLRVLFLSNNRGITNEVINQIKISCPLISHLDISSCLSINQFVIP ILCKMQSLEYIDISFDVMINEDGIISLVNSLTRLSTFCMQGIAFKQSYLFTEEAQWLNS LKLNFIPMCGDDVMRNIASYCPLLTSVELRMCSGVTDNGIEMLLQKCTKISHLVLGGTS ISQFKMMELVGRGLFIA EHI_087890, 484 aa MSSDKQHYNKKNMGIKQFPKDCISQASIITSIDLSNNEIINLPKEMGPFKVLTHFRMMA NKVSSLPLSFTTLTNLRHLDLNANCFTEFPTQITSLTNLEEIQMIQNQLTSIPDCIGNL VKLQRISFTANFLKSLPKGLAKCVDMNYIELTSNEFEEFPDVICELRKVTILMLQQNRI KEVPDSISKLEKLSGLYLSSNNFGKFPESVCTIPSLTQLELDNNNFVDIPDSLSQLTKL KTLIINKSFISCLNSVDMMSNLCQIVLSDTKCMFLPDLSQNSKLTSLNVIRGYLNEVKS LPPNCSCRFSNNQIESIELPENGVLQYMILSNNRLKVSPNLSMLSKISRLDISQNRITR FNENTCHPTLQQLDISCNPLVEFPVCITKCQSLKILNLSDCHLYDIPSNVLSSLSNLET LYIGCNHLSSLESLSVLKKLRALYLQSNNLLHFPQSIFDLITLKTLFVSNNYITTIPNE ISQLTQLEQLAP

EHI_154170, 253 aa MKIGYNEIMIISKYFDDINDFINLEMRVKRFQGNMERFHFNPIP LNQYSRKLFPNIETFLIYNEKDEIFEDGRINNYVIWHKVKYRVIYKGTIPPEVTSLGVE CFSNYKDLITFRIPTIINKLGVSCFHGCNEERIFVNETILISIEIPRNPEIINRKNIEK KNINAPIIPTTISKLGDYRFEDCTT LRLIIIPTSVSELGEYCFINCTS LTSIDIPTTVSEIGNGCFDGCFRKKNEKEQNVKENI LSKILYK EHI_137910, 1065 aa MFIILYFFIYNCFGKCELNFTSASRYGLNGLIVISSNTRECSGDIKIPNSGIDPISNEE KRVEVIGWNAFSNS LIKSIEIPTSIKIICLNAFSNCNN LQNINIPKSVYRIDYQAFSNCKS LVSINFNNAEIKELGNELFIGCNK LKSIDLSTFSIETIYEDTFNGCSN LINVILPYTLREIKESAFSGCTS LTTINLHNTLINSIKEKAFKKCTS LITITFPSTLTSLSNNSFEKCTSLTSIIFQSVLSSSNNNIPFNECNKLKSIYYTGTDLF TNCIFSNNPIQNIYVNSEYQQDFICSIKVIKNINHSPIIYLCKYSDSNECVSYLSYRKS SINQLTNNEICYQRGSTNQKIIINDNKYYLVDCNNNTKIQYYQNKEINNNIISNFIFSV GYKEINDNNIHILREYFISKRIDNIQWITNQCTVTKHLYLNENENSSIKTKDIIIEIYR NGYILNNGIFYIEKNIKMNEIQTLYFGNEQMYREVLINDYFESMNQENEYIFECISKDL CKTYKFNTTNNLFIEDIDFPLLKKEIKTNILAIGYEYFHFYYSPYFSQQVKFTNYCENG KIFRTIGMELQLMSNFVNENECPKDDYSSFSIEIESKNETIEINSRIYKISVLDNRISD STPIYLKFTNSYLRVIPNKCFQITNSSSLLFIPGIYGDKNYIFLKEYNSTNNCNGIYGI SEFSPDQYSYTKFNINDDYSIISYFDNDYKDCGINKIEEASLIRYVLPDINKCQFKSTY TINEFNLIETKYDNSDCKGNIIIQESYALKRCINDTNQLHIIIATFPIKSEDCNNKFKG CINCDLDECYECPSGYGLSMKKDKCIDCSIFTNDCIQCNNQTCTKCNSIDNEIRVYHNN NCLTCKEAFNDNGCLECNVNNCTLCSSINGTLRYYNNGSCKTCEEAYDENCSDCDGIEC KKCKIEYGLKIGRHGCMKCFDLDSKCLECGEGICTKCYNNLLFKNNKCQSCDDIYGKDC LSCDINNCLTCDNNKVVIESKCISCNKLFDHCVHCNTTHCLDCDSIAYSLSNGKCINQN NEQGNNSCLISILLVFSIILFFLI EHI_017710, 426 aa MSDHQIEKKP LTNLTLRQYVFKATEPIAVEVPEELTNLEEMEFFSCVN LDKLILSPHMLTIPSGLCWGCVMLTEVILPSCLRSIEDNAFKSCIR LKTISLPPTITHLGSNSFNDCES LQKITLSSIKKISTR-AFCGCFS LKEVELPECLESIDESAFALCRS LDKISIPNNVSSIGKMAFNECSS LTAISLPQNLQLFDVSALMRCNKLQRIILHGKNEINFKVSYHVSQLLEENGITCNNIEL INGDDLIFSCNEIPSSISSIGDSYYKECEFKTTSFIIPSTITSLGESAFCLCGN

LKKIVISSSITNLPDYCFAYCTD LIEIKIPTSITSIGTRCFHNCIS LKEIYVPDSVIHIGESCFSNCTK LTSIKIPTTCIEKENLSQVFLGSKKLSSINGNSFDYTDECLCDEFIEEDDNDEIHAQGW

EHI_123820 500 aa MQLGYNEIMIVSKYFEDINDFINLEIGIKRFRGNMERFHFNPIPLNEYSRKLFPNIETF HIYNGYDKIFEDGKIFKYVIWYDVSYSRYLEEKKEKNEYKNIEYTIYDREEYGNTIPIE VNSLGINCFYRCNDIQSINIPTSVSKIGNECFYQCTS LTSINIPTSISEIGDKCFCGCYS LRTIDIPISISKIGYCCFSGCSS LQTINIPTNVSKIEYWCFKYCTS LKSINIPTSISEIGNGCFEECSS LTSINIEDVKYISEERIFMNEPV LISIEIPKNLKMINGKHIEKKDINEFIIPSSITEIGDYCFKECLS LKSIDIPISISKIGYWCFCECSS LTSINIPTNVSKIGDYCFYGCYS LRTIDIPTNVSKIGYCCFSRCSS LKSIDIPTSVTEIGYWCFCECSS LRTINIPTSVSEIGNYCFYGCSS LTSINIPTNVSKIGGGCFYKCTS LISINIPSSITSFRNGCFYGCGCEEELKKNKSIPEYCFEEYFYEEIR

EHI_128460, 237 aa MQLGCNEIKIVSKYFKDINDFINLEMGVKRFRGNMERFHFNPIPLNQYSRKLFPNIETF HIYNKEDEIFKDGKIFKYVIWYKVNYSKYLKEKEEGNICKNIEYTKEDREEYGNTIPIE VNSLGNRCFRWCGDIQSINIPTNVNELGNGCFYECSS LTSINLPPSVSKIGDECFYRCSS LTSIKIPTSVGKIGEYCFYDCSS LKSIEIPPRVNKIREGCFCNCTSLTSVTMTSSVTSF

EHI_176480, 241 aa MQLGYNEIMIVSKYFEDINDFINLEMGVKRFEGNMERFHFNPIPLNQYSIEFFPNIETF HIYNEEDEVFEGGRIFKYVIWYKVSYSRYLEEKEKGNVCKNIEYTEEDRNTYGNTIPIE VNSLGDLCFYGCKS LQSINIPTSVSKIGNRCFKYCST LTSINIPTSISKIEDYCFEYCKS LTSIKIPTSVGKIGNKCFSFCTS LTSINIQSSITSFGKGCFYRCGCEEELKKNKQIPKYCFEK

EHI_134140, 439 aa MALGYNEIMIVSKYFEDINDFINLEMGVKRFRGNMERFHFNPIPLNQYSIEFFPNIETF HIYNEYDEIFEDGRIFKYVIWYDVSYSRYLKEKEEMNEYKNIAYTEEDRIKYGNTIPIE VNSLGINCFGQCDIQSINIPTNVSEIGDECFYRCTS LTTINISTSVSEIGNKCFYGCLS LTTINIPTSVIEIGNKCFYECKIQSITIPTSVNEIGNECFKYCKS LTSIEIPTSVSKIGDGCFEYCTS LTTINIPTSVNEIGNECFKYCKS LTSIEIPTSVIEIGNECFKYCKS LTSIEIPTSVSKIGNECFSFCSS LTSIEIPTSVSKIGNKVFYGCDDIQSINIPTSVIEIGNECFKYCKSLTSIEIPTSVSKI GDGCFEYCTSLTSITIPTSVNEIGNE CFKYCKSLTSIEIPTSVIEIGNECFKYCKSLTSIEIPTS

Only with Salmonella enterica EHI_119470, 574 aa MKHTLNIDSIQQVALYFETIDDYKNIILSSKKFQHLLEYFKSNPIPLNDETIKFFPNIN TYNLFSLQDTFYESLSSLKICFYCRISYQRSLKEIKKGNTCKFIYYSKEDKKKFGDILP SQVKKIGKECYCNSPS LSIFEVPYSIVEIGDKSFMNCTS LTSIVLSSSITRLCNYTFHKCIS LKKIKLPQSLKHIGYNCFSGCSS LNSIQLPEMLLSIGDYCFDFCVN LTSLAIPSSITYFGIQCFCNCSSLKSLSIPSNLTSFNRQQFYGCSSLEEINVPAPYWIV GKHLFFNKMKLVGCLPSTINSINKIPVAFDMNQTSFEVPTEVKSFGNKCFQDEHS LRSISIKKFIDRLDNYCFEGCLLVTSISLPSTIKVIENGCFCGCIS LKEINIPSSLTRLGDYCFYRCSSLSAVSLPSSLKVIPYQCFSGCSS LTSIDLPQGVLKLSDQSFKDCAT LQSINIPSTITKIEVGCFNNCLRLSEVSFYSPIQKIPDFCFSNC LQTITLPLSVISIGNRSFCNCVK LQSIQIPSLTTSLGNDCFTNCSSLVSLGSLPKNAILGDDCFFGCEKLFY

EHI_127100, 482 aa MSSEESITLSSCSSTSTAYSEQIQLAIAETDTFGFFSFEVDPLFLVAQYLQTESDFINF IQISKRFQVVLSYFQSNPISV NSLRLFPQIKTQYLFNKNDKRIKGVSKYVYRCLLTHTEAKKLNRSNVSFKRVVFKKDKN AKKHHIPKLTTDLADCCFTKSVY LKSILIPSTVTLLGSDCFLDCSS LSKLVLCSSLIHLGISSFENCKS LRHVVLPLLIKEIPDNCFKECTQ LKDVVLPSCITRIGHSSFENCLS LTEIINTSNIIELGQEAFKGCHS

LKQLHFSSSLNYIGVSCFDDCDL VKINPPPLSSLKCITNKQSLDNKTPSSNHIVLTSDDCIEIMEEIPLTINVIGRSAFKYI MPSQLILPNSITRIESNAFYQCTL LTSIIFSTSLKVIESRAFMGCSH LKSISLPASIQFIGDEAFNNCYE LTSIVVEGFPCIG-YHAFSHCNELQNIIIPPFSNQNTNDIFFGIKIPSK PCCIV EHI_112290, 213 aa MENTTKRNDKERMRLGYNEIMIVSKYFEYINDFINLEEREKKETNECKNIEYTQEDRNT YGTTIPIEVNSLGDICFRWCDDIQSINIPTNVSKIGDKYFEYCTSLRIINIPTNVSKIG DKYFERCSSLTSNSYKNIQFVGEDRIIVNEQIMASIPIGRLKIINEKEIKPKNINEFVI PTTVTKLGNYCFENCYS LTSIKIPTSVSEIGKECFL EHI_041470, 448 aa MQLGYNEIMIVSKYFEDVNDFINLEMGIKRFQGNMERFHFNPIPLNQYSKLFPNIETFH IYNKYDEIFKDGKIIKYVIWYKVEYSRYLKEKEEMNECKNIEYTEEDRKKYGNTIPIEV NSLGINCFYGCNDIQSINIPINVSKIGECCFLCSS LKSINIPTSVSKIGDSCFSGCLS LTSITVPPSVTKIGYGCFDGCSSTSINIPTSVSEIGYGCFDGCSSLTSINIEDIKYISE ERIFMNEPVLISIEIPKNLKMINGKNIEKKDINKFIIPSTITKLGYECFEGCSSLI LINI--PTSVE—IGNECFYGCES LKSIEIQTNVTKIGNECFEYCKS LTSINIPTSVIEIGNC-FRECTS LTSIEIPTSVNEIRNTCFGWCESLILINIPTSISEIGEYCFYECSS LTSINIPTSFTSFEICCFYHCGCEEKLKKNKKIPKYCFKKY _______________________________________________________________________________________________